This protocol describes one-step real time quantitivie PCR to quantify relative levels of a particular mRNA sequence between two samples. This method describes use of a flourescent labeled probe (FAM).

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This protocol describes one-step real time quantitative reverse transcription PCR to quantify relative levels of a particular mRNA sequence between two samples. This technique is also called known commerically as Taqman, qRT-PCR, real-time PCR. This particular protocol describes use of a flourescent labeled probe (FAM) to provide readings.

== Starting Materials ==

== Starting Materials ==

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* Validated PCR primers & probe that are efficient over the range of RNA that you are assaying.

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* Validated PCR primers & probe that are efficient over the range of RNA that you are assaying. Primers are typically designed with a 58 degree Tm but may vary by lab.

* Use a master mix to ensure a consistent amount of enzyme in each tube that you will be comparing.

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== Protocol Endorsements ==

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[[Category:Protocol]]

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[[Category:In vitro]]

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[[Category:RNA]]

Latest revision as of 12:43, 9 October 2007

This protocol describes one-step real time quantitative reverse transcription PCR to quantify relative levels of a particular mRNA sequence between two samples. This technique is also called known commerically as Taqman, qRT-PCR, real-time PCR. This particular protocol describes use of a flourescent labeled probe (FAM) to provide readings.